Characterization of depth-related changes and site-specific differences of microbial communities in marine sediments using quinone profiles

ABSTRACT:   Respiratory quinone compositions were analyzed by a high-performance liquid chromatography to characterize the depth-related changes and site-specific differences of microbial communities in marine sediments. Two deep-sea sediment samples and one coastal sediment sample were investigated from three sites on the coast of Japan, Sagami Bay, Suruga Bay and Tokyo Bay. Although depth-related changes in microbial community structures were observed, site-specific differences appeared to have greater influence on overall community structures. A variety of quinone homologs was commonly identified at all sampling sites and depths examined, but a few minor quinone fractions, mainly derived from Actinobacteria , were detected only at specific sampling sites. Methylmenaquinone-7 (MMK-7) was the major component throughout the examined depths in Sagami Bay and Tokyo Bay. Most abundant quinone homologs changed with each depth in Suruga Bay. Menaquinone-6 (MK-6) predominated in the 0 to 2-cm layer (19%). Below 0–2 cm, the most abundant homolog in each sampling depth was phylloquinone (K₁; 13%, 4–6 cm), MK-8 (19%, 8–10 cm) and MK-7 (13%, 16–18 cm). The microbial respiratory quinone profiling method shown here, successfully demonstrates the usefulness of this approach to characterize microbial communities in marine sediments.     

Selecting the desirable method for predicting 305‐day lactation yields in Mongolia

In Mongolia, milk production must be improved both quantitatively and qualitatively, and we must find the most desirable method for predicting 305‐day milk yields for cows in Mongolia. Therefore, the predicted 305‐day milk yield from the test interval method (TIM), multiple‐trait prediction (MTP) and the random regression model (RRM) were compared. The daily milk production records during 305 days for cows calving from 1986 to 2007 from the National Livestock Breeding Center in Japan, and the test day records every month for cows calving from 1985 to 2005 from the Hokkaido Dairy Milk Recording and Testing Association were used. Wilmink's model for the average of lactation curve in MTP was adopted. A cubic Legendre polynomials and the parameters of Wilmink's function were used for RRM. The results suggested that RRM was the most desirable method for predicting 305‐day milk yields in Mongolia because estimates by RRM were the most accurate when using over four records. Further analyses will be required to examine the properties of RRM when predicting 305‐day milk yields using test day records in Mongolia.     

Differential cytokine gene expression in CD4+ and CD8+ T cell subsets of calves

The distinct patterns of cytokine expression in CD4+ and CD8+ T cells are well understood in mice and humans. However, little information is available about cytokine expression in bovine CD4+ and CD8+ T cells. In this study, mRNA expression of 19 different cytokines was analyzed in CD4+ and CD8+ T cells of calves with or without Concanavalin A (Con A) stimulation. CD4+ and CD8+ T cell populations were enriched to 98% purity by positive selection using magnetic cell sorting (MACS). CD4+ T cells spontaneously expressed the mRNAs of interleukin-1alpha (IL-1alpha), IL-1beta, IL-2, IL-6, IL-7, IL-8, IL-10, IL-18, IFN-gamma, TNF-alpha, TNF-beta and TGF-beta, and augmented the mRNA expression of IL-10, IFN-gamma and TNF-beta after Con A stimulation. The mRNAs of IL-3, IL-4, IL-5, IL-13 and GM-CSF were newly expressed in Con A-stimulated CD4+ T cells. CD8+ T cells displayed spontaneous mRNA expression of IL-6, IL-18, TNF-alpha, TNF-beta and TGF-beta, and newly expressed the mRNA of IL-2, IL-7, interferon-gamma (IFN-gamma) and GM-CSF after Con A stimulation. It was found that CD4+ T cells expressed the mRNA of 17 cytokines except for IL-12 and IL-15, while CD8+ T cells expressed only the mRNA of 9 cytokines after Con A stimulation. The profile of cytokine mRNA expression was substantially different in the CD4+ and CD8+ T cells of calves, indicating that CD4+ T cells can be distinguished from CD8+ T cells by the cytokine gene expression of IL-1alpha, IL-1beta, IL-3, IL-4, IL-5, IL-8, IL-10 and IL-13. Differential cytokine expression between CD4+ and CD8+ T cells serve to interpret an individual function of T cell subsets in the immune system of calves.     

Staining patterns for actin and villin distinguish M cells in bovine follicle-associated epithelium

M cells play a central role in the initiation of mucosal immune responses. However, a primary source of difficulty for investigations of this is the lack of an available specific marker for bovine M cells. As M cells possess irregular and short microvilli, we investigated the distribution and localization of the microvillar proteins actin and villin by immunohistochemistry of the gut of calves. In ileum of the calf, actin and villin were clearly and continuously immunostained in the brush border of the villous epithelia, however, discontinuous immunostaining with patches of no staining were observed in follicle-associated epithelium (FAE). Electron microscopy revealed that M cells had irregular microvilli and lacked the typical brush border, and it was inferred that these patches of no staining might be the intercellular crevices of M cells. As the microvilli of M cells were very sparse, there were several areas of weak immunostaining in calf jejunal FAE. These results suggest that M cells in calf FAE are detectable by the absence of staining for actin and villin.     

Development of a multiplex polymerase chain reaction method for simultaneous detection of eight events of genetically modified maize

In this study, we developed a novel multiplex polymerase chain reaction (PCR) method for simultaneous detection of up to eight events of genetically modified (GM) maize within a single reaction. The eight detection primer pairs designed to be construct specific for eight respective GM events (i.e., Bt11, Event176, GA21, MON810, MON863, NK603, T25, and TC1507) and a primer pair for an endogenous reference gene, ssIIb, were included in the nonaplex(9plex) PCR system, and its amplified products could be distinguished by agarose gel and capillary electrophoreses based on their different lengths. The optimal condition enabled us to reliably amplify two fragments corresponding to a construct specific sequence and a taxon specific ssIIb in each of the eight events of GM maize and all of nine fragments in a simulated GM mixture containing as little as 0.25% (w/w) each of eight events of GM maize. These results indicate that this multiplex PCR method could be an effective qualitative detection method for screening GM maize.     

Cucumber mosaic virus isolated from Aconitum spp. in Japan

Viruses were isolated from leaves of plants of Aconitum species with symptoms such as mottling and yellowing in Hokkaido and Gunma prefectures in Japan. These viruses were identified as Cucumber mosaic virus (subgroup II) based on particle morphology, host range, aphid transmission, and serology.     

Assemblages of vertical migratory mesopelagic fish in the transitional region of the western North Pacific

Assemblages of vertical migratory mesopelagic fish are described for the nighttime upper 20 m layer of the transitional and adjacent waters in the western North Pacific. Twenty-three mesopelagic fish species belonging to six families and 16 genera were collected during June 1996. Family Myctophidae was most speciose, represented by 17 species, followed by Sternoptychidae (two spp.) and Bathylagidae (one spp.). Six assemblages are recognized based on their species composition: Subarctic, Northern Transition, Southern Transition, Subtropical, Slope Water and Seamount Assemblages. These assemblages are also characterized by size composition and biomass of the catches. The distributions of the Subarctic, Northern Transition, Southern Transition and Subtropical Assemblages are approximately defined by the Kuroshio and Oyashio fronts and the Subarctic Boundary. Slope Water and Seamount Assemblages are closely related to the slope water around the shelf edge off Japan and waters around the Shatsky Rise and the Emperor Seamount chain, respectively. The importance of the Southern Transition Assemblage area between the Kuroshio front and Subarctic Boundary as spawning and nursery grounds of many subarctic, transitional and subtropical fishes is elucidated, based on the geographical distribution patterns of their larvae and juveniles.     

Age-related changes in leukocytes and T cell subsets in peripheral blood of Japanese Black cattle

It is well known that the immune system changes with age during development and maturation in Holstein cattle. But age-related changes in leukocytes and T cell subsets in peripheral blood of Japanese Black cattle still remain unclear. The aim of the present study was to investigate comparative changes of leukocytes (granulocytes, monocytes, B cells and T cells) and T cell subsets (CD4⁺, CD8⁺, γδ, CD8⁺γδ and WC1⁺γδ T cells) in Japanese Black cattle aged 0.5, 1, 2, 6, 18 and 36–41 (adult) months on flow cytometry using specific monoclonal antibodies for the cell surface markers. T cell proportion was approximately 40% in 2-month-old cattle and decreased to 20.6% in adults. In contrast, B cell proportion significantly increased from 7.4% to 28.2% with age. In T cell subsets the percentage of CD4⁺ T cells significantly increased from 40.5% to 60%, but that of WC1⁺γδ T cell subset significantly decreased with age. The percentages of CD8⁺ and CD8⁺γδ T cells did not change. The present study details the proportional changes in leukocyte and T cell subsets with age in the peripheral blood of Japanese Black cattle and these findings are similar to those described for Holstein cattle.     

Central insulin suppresses feeding behavior via melanocortins in chicks

Growing evidence suggests that insulin interacts with both orexigenic and anorexigenic peptides in the brain for the control of feeding behavior in mammals. However, the action of central insulin in chicks has not yet been identified. In the present study, we investigated the effects of central injection of insulin on feeding behavior in chicks. Intracerebroventricular (ICV) administration of insulin, at doses that do not influence peripheral glucose levels, significantly inhibited food intake in chicks. Central injection of insulin in chicks significantly increased expression of pro-opiomelanocortin (POMC) mRNA, and decreased that of neuropeptide Y (NPY) mRNA. Finally, co-injection of the melanocortin antagonist (SHU9119 or HS014) prevented the reduction in food intake caused by ICV administration of insulin. These data suggest that insulin functions in chicks as an appetite-suppressive peptide in the central nervous system, and that the central melanocortin system mediates this anorexic effect of insulin, as in mammals.